Invitrogen™ LIVE/DEAD™ Viability/Cytotoxicity Kit, for mammalian cells

Catalog No.	Quantity
L3224	1 kit

Catalog No. L3224 Supplier Invitrogen™ Supplier No. L3224

Description

Useful with a variety of fluorescence-detection methodologies

The LIVE/DEAD Viability/Cytotoxicity Kit is a quick and easy two-color assay to determine viability of cells in a population based on plasma membrane integrity and esterase activity. The kit can be used in flow cytometry, fluorescence microscopy, and with fluorescence microplate readers.

• Faster, Safer, more sensitive
• Less expensive

Easily Use with a Wide Variety of Techniques and Cell Types
Ubiquitous intracellular esterase activity and an intact plasma membrane are distinguishing characteristics of live cells. The LIVE/DEAD Viability/Cytotoxicity Kit quickly discriminates live from dead cells by simultaneously staining with green-fluorescent calcein-AM to indicate intracellular esterase activity and red-fluorescent ethidium homodimer-1 to indicate loss of plasma membrane integrity. It is adaptable to most eukaryotic cells where cytotoxic conditions produce these cellular effects. The assay is useful with a variety of fluorescence-detection methodologies.

Sensitive, Safe, and Efficient
The LIVE/DEAD Viability/Cytotoxicity Kit is more sensitive than Trypan blue exclusion, a commonly used method for live/dead cell discrimination. The cost-effective LIVE/DEAD Viability/Cytotoxicity Kit is highly sensitive due to the bright fluorescence of both dyes upon interacting with either live (for calcein-AM) or dead (for ethidium homodimer-1) cells. Background levels are low due to the fact that both dyes are virtually non-fluorescent prior to interacting with cells.

LIVE/DEAD Assays Available for a Broad Range of Applications
A selection of Invitrogen LIVE/DEAD Viability Assays is offered for mammalian cells, bacteria, yeast and fungi, as well as Fixable Dead Cell Stain Kits for use in intracellular staining for flow cytometry. All LIVE/DEAD assays provide quick, positive discrimination between viable and non-viable cells.

Cell Analysis, Cell Counting and Viability, Cell Counting, Viability, and Cryopreservation, Cell Culture, Cell Metabolism, Cell Viability and Cytotoxicity, Cell Viability, Proliferation and Function, Cellular Imaging, Cellular Toxicology Assays, Drug Discovery and Development, Flow Cytometry, Flow Cytometry Viability and Cytotoxicity Assays, Flow Cytometry of Cellular Processes, High-Content Screening (HCS), Immunofluorescence (IF), Immunofluorescence Staining and Detection, Target-Based ADME/Tox Assays

Order Info

Shipping Condition: Room temperature

Specifications

• Cell Type: Mammalian Cells
• Color: Green, Red
• Content And Storage: Store in freezer at -5°C to -30°C and protect from light.
• Description: LIVE/DEAD™ Viability/Cytotoxicity Kit, for mammalian cells
• Detection Method: Fluorescence
• For Use With (Application): Viability Assay
• For Use With (Equipment): Fluorescence Microscope, Flow Cytometer, Microplate Reader
• Product Type: Viability/Cytotoxicity Kit
• Dye Type: Other Label(s) or Dye(s)
• Emission: 517/617
• Excitation Wavelength Range: 494, 528 nm
• Format: Tube(s), 96-well plate, Slide(s)
• Product Line: LIVE/DEAD
• Quantity: 1 kit
• Shipping Condition: Room Temperature

Frequently Asked Questions (FAQs)

I need to use a dead cell control for my viability assay. Do you have a protocol for killing cells for this?

Heat killing is commonly used. Place your cells in a tube in buffer and heat at 60^oC for 20 minutes. You can also kill your cells by fixing them with ice cold 70% ethanol for 15 minutes. The ethanol-killed cells can then be stored at -20^oC until needed, at which point you wash out the ethanol and replace with buffer.

I am doing a Live/Dead assay using Calcein, AM, for live cells and ethidium homodimer-1 for dead cells. Can I fix the cells after labeling and retain the staining?

This is not recommended. Neither Calcein nor ethidium homodimer-1 bind to any cellular components upon fixation. There is no guarantee that the dyes will be retained upon fixation or any subsequent wash steps. We recommend scoring for live and dead cells as soon as possible after staining.

How should I store the LIVE/DEAD Viability/Cytotoxicity Kit, for mammalian cells (Cat. No. L3224)?

We recommend storing the LIVE/DEAD Viability/Cytotoxicity Kit, for mammalian cells (Cat. No. L3224) in the freezer at -5 degrees C to -30 degrees C and protected from light.

What fluorescent viability assays can I use on the Countess II FL automated cell counter?

We have validated the following kits for use on the Countess II FL Automated Cell Counter:

LIVE/DEAD Viabilty/Cytoxicity Kit (Cat. No. L3224) containing calcein AM and ethidium homodimer-1
ReadyProbes Cell Viability Imaging Kit, Blue/Green (Cat. No. R37609)
ReadyProbes Cell Viability Imaging Kit, Blue/Red (Cat. No. R37610) containing NucBlue Live/NucGreen Dead and NucBlue Live/propidium iodide
See this Application Note for details - https://www.thermofisher.com/content/dam/LifeTech/global/life-sciences/Laboratory%20Instruments/Files/0415/CO014723-Countess-II-FL-Viability-Appnote_FHR.pdf.

How do I prepare dead cell controls for LIVE/DEAD cell viability assays?

There are two easy options. One is to heat-inactivate the cells by placing at 60 degrees C for 20 minutes. The second is to subject the cells to 70% ethanol. Alcohol-fixed cells can be stored indefinitely in the freezer until use, potentially up to several years.

Centrifuge cells, pellet, and remove supernatant.
Fix cells: Add 10 mL ice cold 70% ETOH to a 15 mL tube containing the cell pellet, adding dropwise at first while vortexing, mix well.
Store in freezer until use.
When ready to use, wash twice and resuspend in buffer of choice.

For Research Use Only. Not for use in diagnostic procedures.